Detection assay for protein-polynucleotide conjugates
AssigneeAmgen Inc.
InventorsMai Thayer, Sara Humphreys
Life of this patent
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Abstract
The present invention relates to methods for detecting and quantifying intact protein-polynucleotide conjugate molecules in various sample matrices. In particular, the methods utilize triplex forming oligonucleotides in combination with protein-specific binding partners to respectively detect the polynucleotide and protein components of the conjugate molecules.
Claims37
An assay system for detecting a protein-polynucleotide conjugate molecule in a sample, comprising:
- (a) a triplex forming oligonucleotide (TFO) that is covalently linked to a tag, wherein the TFO is capable of hybridizing to the polynucleotide in the conjugate molecule;
- (b) a surface comprising a capture reagent that specifically binds to the tag covalently linked to the TFO;
- and
- (c) a detection reagent comprising a detectable label coupled to a binding partner that specifically binds to the protein in the conjugate molecule.
An assay system for detecting a protein-polynucleotide conjugate molecule in a sample, comprising:
- (a) a TFO that is covalently linked to a tag, wherein the TFO is capable of hybridizing to the polynucleotide in the conjugate molecule;
- (b) a surface comprising a capture reagent that specifically binds to the protein in the conjugate molecule;
- and
- (c) a detection reagent comprising a detectable label coupled to a binding partner that specifically binds to the tag covalently linked to the TFO.
A method for detecting antibody-polynucleotide conjugate molecules in a biological sample comprising:
- (a) contacting the sample with a TFO that is covalently linked to a tag under conditions that allow the TFO to hybridize to the polynucleotide in the conjugate molecule, thereby forming a hybridization mixture;
- (b) contacting the hybridization mixture with a surface comprising a capture reagent that specifically binds to the tag covalently linked to the TFO;
- (c) contacting the surface with a detection reagent, wherein the detection reagent comprises a detectable label coupled to a binding partner that specifically binds to the antibody in the conjugate molecule;
- and
- (d) detecting a signal from the detectable label.
Dependencies are inferred from the claim text.